Therapeutic agent and method for feline AIDS virus infections and feline atopic dermatitis

ABSTRACT

A therapeutic agent for feline immunodeficiency virus (FIV) infections, (including the treatment of the anemia and chronic stomatitis caused by infection with a FIV) comprising a feline interferon preparation containing a feline interferon as a principal agent, and a therapeutic method for FIV infections comprising administering a feline interferon preparation containing a feline interferon as a principal agent to a cat every day are disclosed. Furthermore, a therapeutic method and agent for feline atopic dermatitis are disclosed. The preferred feline interferon, is an ω-feline interferon.

Therapeutic agent and method for feline AIDS virus infections and felineatopic dermatitis.

1. Technical Field

The present invention relates to a therapeutic agent and method forfeline AIDS virus (FIV) infections and feline atopic dermatitis. A FIVbelongs to Lentiviridae of Retroviridae, and cats living freely outdoorsare often infected with it.

FIV infections are said to undergo three stages; an initial acute stageof virus infection, latent stage, and lastly, a chronic disease stage ofimmunodeficiency.

In the initial stage of infection, fever and lethargy can be observed,and lymphopenia and neutropenia occur. The skin and digestive tracts canbecome infected with microbes, but these are secondary infections byneutropenia. Vomition and anemia can also be observed. After thesymptoms in the acute stage vanish, the latent stage continues forseveral months to several years.

In the last stage, chronic diseases such as chronic stomatitis, chronicrespiratory organ diseases, anemia, intractable secondary infections andenteritis occur, and the carriers become gradually weak in severalyears, and finally die. Furthermore, opportunistic infections such asHaemobartonella diseases and Cryptococcus diseases can be seen.

2. Background of the Related Art

Therapy of FIV infections is still untapped, and no therapeutic methodhas been developed for eliminating FIV, i.e., a retrovirus from thefeline body after infection. As therapeutic agents for human AIDS aredeveloped, it can be considered to use them as anti-FIV drugs for catsin the future, but the application of a reverse transcriptase inhibitorsuch as AZT (azidothymidine) seems problematic for the time being,considering its side effects versus the desired effect to be achieved.

At present, therapeutic methods for symptoms from secondary infections,such as the administration of antibiotics, infusion and blood infusionand administration of steroid hormone preparations are used.

Anemia as one of FIV infections can be seen in the early stage and laststage of FIV virus infection, and at the onset, vitality vanishes andappetite diminishes or is lost. Furthermore, erythroid values such aserythrocyte number, hemoglobin and hematocrit decrease. Symptomatictherapeutic methods such as the administration of erythropoietin and theinfusion of vitamins, amino acids, etc., and as the case may be, bloodinfusion are used. However, most carriers die, though these methods havesome macrobiotic effect.

Chronic stomatitis as one of FIV infections is an intractable disease,and the inflammation of the gum near the roots of molar teeth causesheavy swelling, not allowing eating. Weakened cats in this stage come tohospital. Steroid hormone preparations such as Depo-Metrol areadministered to temporarily improve the inflammation as a symptomatictherapeutic method. However, the disease recurs in 2 to 3 weeks, and theadministration of steroid hormone preparations is practiced again. Therecurrence period becomes gradually shorter, and partly because of sideeffects by steroid hormone preparations, the cats ultimately die.

As a feline interferon, a recombinant ω-interferon preparation isalready approved as a therapeutic agent for calicivirus infections, andis marketed under a trade name of “INTERCAT” since Feb., 1994. Theinventors studied a therapeutic method for FIV infections using thisrecombinant ω-interferon, and as a result, completed the presentinvention.

Presently known interferons include alpha (α) interferons, beta (β)interferon, gamma (γ) interferon, omega (ω) interferon and tau (τ)interferon. As human interferons, three types of α, β and γ arepractically applied, and as a feline interferon, an ω-interferon only ispractically applied. “INTERCAT” is a recombinant ω-feline interferonpreparation, and it is an injection preparation obtained by infectingBombyx mori with a baculovirus recombined with the gene of an 107-feline interferon, producing the interferon in the Bombyx mori,extracting and purifying it, adding gelatin and D-sorbitol as astabilizer and recipient, and freeze-drying the mixture. The recombinantω-feline interferon is a glycoprotein with a molecular weight of about25,000, and its protein portion has the amino acid sequence as shown inSEQ ID NO:1 of the sequence listing.

The ω-feline interferon can also be produced by other methods than theBombyx mori method. For example, it can be produced by transientexpression methods using animal cells such as simian COS cells and generecombination techniques using Chinese hamster ovary (CHO) cells,Escherichia coli, yeast, transgenic animals, etc.

As for the usage and dose of “INTERCAT” approved as a therapeutic agentfor calicivirus infections, it is specified to administer 2.5˜5 MU/kg offeline interferon intravenously three times every other day. In thiscase, MU (mega unit) is a method for expressing a titer with theantiviral activity of an interferon as an indicator, and expresses onemillion units. The inventors attempted to treat FIV infections,particularly anemia and chronic stomatitis, according to the same usageand dose of every-other-day administration as approved for treatingcalicivirus infections, but the expected effects could not be obtained.So, the inventors continued their studies by changing the usage anddose.

A task of the present invention is to provide a new excellenttherapeutic agent and method for FIV infections.

On the other hand, for feline atopic dermatitis, there is nosatisfactory therapeutic method even for human atopic dermatitis.Symptomatic therapeutic methods using steroid hormone preparations arefrequently adopted. Steroid hormone preparations have side effects, andthe symptomatic therapeutic methods are insufficient in therapeuticeffect. Feline atopic dermatitis is an intractable disease. Generallyobserved feline allergic dermatitis is mostly atopic dermatitisincluding miliary eczema relating to parasites (such as fleas) andeosinophilic granuloma syndrome. Hitherto, drugs such as prednisoloneand amcinolone are said to be effective, and they tend to be used morefrequently. However, these drugs have the problem of side effects.

It was reported in an American medical magazine in 1990 (M. Boouniewwiczet al., American J. Medicine, 8.8, 365-370 (1990)) that a human γ(gamma)interferon is effective for human atopic dermatitis.

However, this method is not sufficient in the effect of treating felineatopic dermatitis since human interferons are different in their actionthan feline interferons.

Another object of the present invention is to provide a new excellenttherapeutic agent and method for feline atopic dermatitis.

Disclosure of the Invention

The inventors studied to achieve the above objectives, and as a result,found a therapeutic method for FIV infections by injecting a therapeuticagent containing a feline ω-interferon into cats.

An effective therapeutic method has been invented, in which the anemiaand chronic stomatitis caused by infection with a FIV (confirmed to bepositive in anti-FIV antibody by virus tests of feline blood) aretreated by using a therapeutic agent containing a feline ω-interferon.

Furthermore, it has been found that a therapeutic agent containing an(ω-feline interferon is a new excellent therapeutic agent for felineatopic dermatitis.

The following objectives of the present invention have been industriallyadvantageously achieved by the present invention with the followingconstitution:

[1] A therapeutic agent for FIV infections, comprising a felineinterferon preparation containing a feline interferon as a principalagent.

[2] The therapeutic agent for FIV infections, wherein the felineinterferon is a a feline ω-interferon.

[3] The therapeutic agent for feline AIDS virus infections, wherein thefeline ω-interferon is a recombinant interferon.

[4] The therapeutic agent for FIV infections, wherein the felineω-interferon is a glycosylated interferon having the amino acid sequenceshown in SEQ ID NO:1.

[5] The therapeutic agent for FIV infections, according to any one of[1] through [4], which is used for treating anemia caused by infectionwith FIV.

[6] The therapeutic agent for FIV infections, according to any one of[1] through [4], which is used for treating chronic stomatitis caused byinfection with FIV.

[7] A therapeutic method for FIV infections, comprising the step ofadministering a feline interferon preparation containing a felineinterferon as a principal agent to a cat continuously every day.

[8] The therapeutic method for FIV infections, wherein the felineinterferon is a feline ω-interferon.

[9] The therapeutic method for FIV infections, wherein the felineω-interferon is a recombinant interferon.

[10] The therapeutic method for FIV infections, wherein the felineω-interferon is a glycosylated interferon having the amino acid sequenceshown in SEQ ID NO:1.

[11] The therapeutic method for FIV infections, according to any one of[7] through

[10], which is used for treating anemia caused by infection with FIV.

[12] The therapeutic method for FIV infections, according to any one of[7] through [10], which is used for treating chronic stomatitis causedby infection with FIV.

[13] The therapeutic method for FIV infections, according to any one of[7] through [12], wherein the feline ω-interferon is administered as adose of 0.5 MU/kg˜2.5 MU/kg per cat body weight once or more per day for5 or more consecutive days.

[14] A therapeutic agent for feline atopic dermatitis, comprising afeline interferon.

[15] The therapeutic agent for feline atopic dermatitis, wherein thefeline interferon is a feline ω-interferon.

[16] The therapeutic agent for feline atopic dermatitis, wherein thefeline ω-interferon is a recombinant interferon.

[17] The therapeutic agent for feline atopic dermatitis, according to[15] or [16], wherein the feline ω-interferon is a glycosylatedinterferon having the amino acid sequence shown by SEQ ID NO:1.

[18] The therapeutic method for feline atopic dermatitis, wherein thetherapeutic agent for atopic dermatitis stated in any one of [14]through [17] is injected into a cat.

[19] The therapeutic method for feline atopic dermatitis, wherein theinjection is a subcutaneous injection.

[20] The therapeutic method for feline atopic dermatitis, wherein theinjection dose is 0.1 -5 MU/kg.

THE BEST EMBODIMENTS OF THE INVENTION

It is preferable that the feline interferon used in the presentinvention is a feline ω-interferon which can be as produced naturally,synthetically synthesized, or produced by any gene recombinationtechnique.

For example, a feline ω-interferon produced by a genetic recombinationand marketed under a trade name of “INTERCAT”(produced by TorayIndustries, Inc.) can be used.

“INTERCAT” has been approved and practically applied as a therapeuticagent for feline calicivirus infections, and mainly contains aglycosylated interferon having the sequence of 170 amino acids shown inSEQ ID NO:1, and it is obtained by infecting larvae of Bombyx mori witha recombinant baculovirus, (i.e., an insect virus recombined with thegene of a feline ω-interferon), and extracting, separating and refiningthe interferon produced in Bombyx mori.

However, the feline ω-interferon of the present invention is notnecessarily limited to recombinant feline interferon.

The ω-feline interferons produced by gene manipulation using bacterialcells (such as Escherichia coli and Bacillus subtilis) animal cells(such as CHO) and also the ω-interferon produced from feline cells canbe used. However, presently, the feline ω-interferon produced fromBombyx mori is available at low cost.

First, the therapeutic method for FIV infections is described. Thetherapeutic method is to inject a therapeutic agent containing a felineω-interferon into a cat once or more per day for 5 or more consecutivedays. The dose of the feline interferon is 0.5 MU/kg˜2.5 MU/kg per catbody weight.

It is practical to administer the therapeutic agent once a day. It canbe administered once or more per day, but it is preferable to administerat least once a day. It is desirable to administer every day, instead ofevery other day, and it is more desirable to administer continuously for5 days or more.

After administering for 5 consecutive days or more, the administrationcan be once suspended, and subsequently administration can be effectedagain for 5 or more consecutive days.

The dose can also be smaller than 0.5 MU/kg, but if the dose is smallerthan 0.5 MU/kg, the therapeutic effect is also weaker. On the contrary,even if the dose is larger than 2.5 MU/kg, the therapeutic cost simplyincreases, without giving any correspondingly higher effect in mostcases.

The injection route can be subcutaneous or intravenous. Intramuscularinjection can also be used. However, subcutaneous injection can bepractically and simply effected. When the therapeutic method of thepresent invention was used for treating the anemia as one of feline AIDSvirus symptoms, recovery from lethargy and increase of appetite could beachieved, and erythroid values such as erythrocyte number, hemoglobinand hematocrit increased.

When used to treat chronic stomatitis, the stomatitis accompanying theulcers and granulomata of fauces improved and appetite and vitality wererestored.

Feline atopic dermatitis it described below. The therapeutic agent forfeline atopic dermatitis of the present invention is a preparationcontaining a feline ω-interferon as a principal agent, and when it isused, a solution obtained by dissolving it into physiologic saltsolution or infusion solution or any other solution is injected. Theinjection route can be subcutaneous, intravenous or in tramuscular.Subcutaneous injection is preferably simple and practical. The number ofadministrations is not especially limited, but it is practical toadminister once a day every day or 1 to 3 times per week. The dose isnot limited either, but is usually 0.1 to 5 MU/kg. A preferable range is1 to 2.5 MU/kg. The administration effect can be clearly observed fromabout the 2nd week in most cases.

The feline ω-interferons usually do not cause remarkable fever afteradministration unlike in human beings, and even if fever occurs, bodytemperature elevation as slight as about 1° C. only occurs for a while.Serious side effects such as vomition and diarrhea do not occur.

EXAMPLES

The present invention is described below in reference to examples, butis not limited thereto or thereby. The blood cell count is in number ofcells per microliter (/μl).

Example 1

A recombinant feline ω-interferon preparation (trade name: INTERCAT) wasadministered to a Japanese cat (female) of 3 to 4 years of age that hadbeen suffering from anemia caused by FIV infection. On the day of thefirst medical examination, the body weight was 3.7 kg, and she had lostappetite from the previous day, remained lethargic and showed palemucous membranes.

1.85 ml tetracycline was subcutaneously injected, and a 500 ml vitaminpreparation was subcutaneously dripped. On the following day, the bodyweight became 4.0 kg. By a virus check, she was positive for anti-FIVantibody and negative for FeLV antigen. Blood examination values wereWBC 12,600, erythrocytes 144,000, hemoglobin 3.7 g/dl, hematocrit 12.3%,mean cell volume (MCV) 85 fl, mean cell hemoglobin concentration (MCHC)30.1 g/dl and thrombocytes 163,000.

INTERCAT was dissolved into physiological salt solution, and theINTERCAT solution was subcutaneously injected by 10 MU/day for 3 days.The dose per body weight was 2.5 MU/kg. A 500 ml/day infusion solution(vitamin preparation) was subcutaneously dripped, and a 1.85 mlantibiotic (tetracycline) was subcutaneously injected.

Prom the 4th day, INTERCAT was decreased to 4 MU/day, and subcutaneouslyinjected for 4 days. The dose per body weight was 0.75 MU/kg.

On the 5th day, appetite was restored a little.

On the 8th day, blood examination values were WBC 10,400, erythrocytes358, hemoglobin 4.9 g/dl, hematocrit 28.5 %, mean cell volume (MCV) 65fl, mean cell hemoglobin concentration (MCHC) 30.2 g/dl and thrombocytes178,000. The body weight was 3.4 kg.

Even after the 85th day, vitality and appetite remained normal. Theblood examination values were WBC 11,400, erythrocytes 971, hemoglobin11.8 g/dl, hematocrit 40.3%, mean cell volume (MCV) 42 fl, mean cellhemoglobin concentration (MCHC) 29.3 g/dl and thrombocytes 198,000. Thebody weight was 3.9 kg.

Example 2

A recombinant feline ω-interferon preparation (trade name: INTERCAT) wasadministered to a Japanese cat (male) of 6 to 7 years old who had beensuffering from anemia caused by infection with FIV. On the day of thefirst medical examination, the body weight was 7.55 kg. Appetite haddeclined from the previous day, and he was lethargic.

By a virus test, he was positive for anti-FIV antibody and negative forFeLV antigen. An anti-inflammatory drug, loxoprofen sodium wasadministered (½ tablet twice a day).

The blood examination values were WBC 3,500, erythrocytes 3,670,000,hemoglobin 5.3 g/dI, hematocrit 18.4%, mean cell volume (MCV) 50 fl,mean cell hemoglobin concentration (MCHC) 31.5 g/dl and thrombocytes105,000.

INTERCAT was dissolved into physiological salt solution, and 10 MU/dayof the INTERCAT solution was subcutaneously injected for 3 days. On the4th day, the dose was decreased to 2.5 MU/day, and the INTERCAT solutionwas subcutaneously injected for further 3 days. From the 7th day, thedose was increased to 10 MU/day, and the INTERCAT solution wassubcutaneously injected for 3 days.

On the 9th day, the blood test values were WBC 10,600, erythrocytes4,020,000, hemoglobin 6.4 g/dl, hematocrit 21.0%, mean cell volume (MCV)52 fl, mean cell hemoglobin concentration (MCHC) 30.5 g/dl andthrombocytes 351,000. Appetite was restored a little. The body weightwas 7.3 kg.

On the 18th day, he came to hospital again due to anorexia. The bodyweight was 7.05 kg. The blood examination values were WBC 6,900,erythrocytes 10,430,000, hemoglobin 16.0 g/dl, hematocrit 51.7%, meancell volume (MCV) 50 fl, mean cell hemoglobin concentration (MCHC) 30.9g/dl and thrombocytes 324,000.

A 10 MU/day solution of INTERCAT was subcutaneously injected for 7 days.An antibiotic (Baytril) was administered (½ tablet twice a day).Infusion was effected on the 18th day only. Even after one month,vitality and appetite remained recovered.

Example 3

A recombinant feline ω-interferon preparation (trade name: INTERCAT) wasadministered to a 10-year-old Japanese cat (male) who had been sufferingfrom chronic stomatitis caused by FIV infection. On the day of the firstmedical examination, the body weight was 4.6 kg, and saliva and theulcers and granulomata of fauces on both sides were observed. By a virustest, he was positive for anti-FIV antibody and negative for FeLVantigen.

INTERCAT was dissolved in physiological salt solution, and 10 MU/day ofthe INTERCAT solution was subcutaneously injected for 3 days. The doseper body weight was 2.17 MU/kg. After the 4th day, the dose wasdecreased to 4 MU/day, and the INTERCAT solution was subcutaneouslyinjected for 4 more days. Only on the 2nd day, a 500 ml infusionsolution (vitamin preparation) was subcutaneously dripped. An antibiotic(Baytril) was administered every day.

On the 3rd day, appetite was restored a little. Saliva was also improveda little. As for the stomatitis, the granuloma on the left side becameslightly less reddish.

On the 4th day, as for the stomatitis, the granuloma on the left sidewas reduced in size and reddishness.

On the 5th day, as for the stomatitis, the granuloma on the right sidevanished, and the granuloma on the left side was further reduced inreddishness.

On the 7th day, appetite was restored, and as for the stomatitis, ulcersand granulomat vanished. On the 10th day, the body weight was 4.95 kg.

Even after six months, the stomatitis was not worsened.

Example 4

A recombinant feline ω-interferon preparation (trade name: INTERCAT) wasadministered to an 8-year-old Japanese cat (male) who had been sufferingfrom chronic stomatitis caused by infection with FIV. On the day of thefirst medical examination, the body weight was 3.4 kg, and saliva andulcers and granulomata of fauces on both sides were observed. By a virustest, he was positive for anti-FIV antibody and negative for FeLVantigen.

INTERCAT was dissolved into physiological salt solution, and 8.5 MU/dayof the INTERCAT solution was subcutaneously injected for 7 days. Thedose per body weight was 2.5 MU/kg.

After the 8th day, the dose was decreased to 4 MU/day, and the INTERCATsolution was subcutaneously injected for 4 more days. On the 2nd dayonly, a 300 ml infusion solution (vitamin preparation) wassubcutaneously dripped. Antibiotics (Dalacin; clindamycin) wereadministered every day.

On the 7th day, appetite was restored. The body weight was 3.75 kg.Saliva improved, and the stomatitis also improved.

After two months, the stomatitis was not especially worsened, but 2.5MU/kg INTERCAT was subcutaneously injected for 7 days.

After six months, the stomatitis was not worsened.

Example 5

A 4-year-old spayed female house cat (short hair, Japanese cat) (white,body weight 2.62 kg) came to the hospital primarily for the main reasonthat red eczema occurred in the abdominal area several days before, andshe was diagnosed with atopic dermatitis. A preparation containing afeline ω-interferon (recombinant) as a principal agent, i.e., “INTERCAT”was dissolved into physiological solution, and a 5 MU/ead (1.9 MU/kg) ofthe INTERCAT solution was subcutaneously injected. The administration of“INTERCAT” was continued at intervals of twice a week. After 8 weeks,the eczema perfectly vanished.

Example 6

A 3-year-old female house cat (Cornish Rex) (white, body weight 2.76 kg)had eosinophilic plaques formed with the hypertrophy of the dorsolumbarskin since several months before, and was cured temporarily by periodicadministration of a steroid hormone preparation. However, after a while,many plaques occurred on the face and the back. On the auricles,portions showing the presence of a fungus existed. The disease wasdiagnosed as atopic dermatitis. A preparation containing a feline107-interferon (recombinant) as a principal agent, i.e., “INTERCAT” wasdissolved into physiological salt solution, and 5 MU/head (1.81 MU/kg)of the INTERCAT solution was subcutaneously injected. An antihistaminicagent previously used was also used. The administration of “INTERCAT”was continued at intervals of once a week. From the 2nd week, thereddishness of the eczema began to vanish, and after 3 months, only atrace remained, showing an almost perfect healing.

Example 7

A 6-year-old spayed female house cat (short hair, Japanese cat)(blackish tiger color, body weight 4.55 kg) had had miliary eczema onthe back due to flea allergy since two years before, and theadministration of a steroid hormone preparation and thorough fleaextermination brought about a lesion. However, since about one yearbefore, many eosinophilic plaques were formed in the abdominal area, andalthough the administration of the steroid hormone preparation showed aneffect, the effect gradually diminished. The disease was diagnosed asatopic dermatitis. A preparation containing a feline ω-interferon(recombinant) as a principal agent, i.e., “INTERCAT” was dissolved intophysiological salt solution, and 5 MU/head (1. 1 MU/kg) of the INTERCATsolution was subcutaneously injected. The administration of “INTERCAT”continued at intervals of once a week. After 2 weeks, reddishness almostvanished, and even after 2 months, the disease did not recur.

Industrial Applicability

The present invention is an effective therapeutic agent containing afeline ω-interferon, for treating anemia and chronic stomatitis causedby FIV infection (confirmed by a virus test of feline blood showinganti-FIV antibody), and also is an effective therapeutic method usingsaid therapeutic agent. Furthermore, the therapeutic agent containing afeline ω-interferon is a new excellent therapeutic agent and method forfeline atopic dermatitis. The present invention is highly industriallyuseful.

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What is claimed is:
 1. A method for ameliorating symptoms associatedwith FIV infections, comprising administering a preparation containingan effective amount of a feline ω-interferon as a principal agent to acat infected with FIV for a time and under conditions sufficient toameliorate symptoms associated with FIV infections.
 2. The methodaccording to claim 1, wherein said feline ω-interferon is a recombinantω-interferon.
 3. The method according to claim 1, wherein said felineω-interferon is a glycosylated interferon having the amino acid sequenceshown in SEQ ID NO:1.
 4. The method a according to claim 1, wherein saidω-interferon is administered for treating anemia caused by infectionwith FIV.
 5. The method according to claim 1, wherein said ω-interferonis administered for treating chronic stomatitis caused by infection withFIV.
 6. The method according to claim 1, wherein said felineω-interferon is administered at a dose of 0.5 MU/kg˜2.5 MU/kg per catbody weight, at least once per day for at least 5 consecutive days.
 7. Athereapeutic method for ameliorating symptoms associated with felineatopic dermatitis comprising administering an effective amount of afeline ω-interferon to a cat suffering from atopic dermatitis for a timeand under conditions sufficient to ameliorate symptoms associated withfeline atopic dermatitis.
 8. The method according to claim 7 whereinsaid feline ω-interferon is administered by subcutaneous injection. 9.The method according to claim 7 wherein said feline ω-interferon isadministered at a dose of 0.1˜5 MU/kg.